CZB and CZB-HEPES Media (1×)

Chemical reagent	Sigma-Aldrich catalog no.	Molecular weight	Final concentration (mm)	Final concentration (g/L)
NaCl	S5886	58.450	81.62	4.76
KCl	P5405	74.557	4.83	0.36
KH₂PO₄	P5655	136.091	1.18	0.16
MgSO₄ · 7H₂O	M1880	246.470	1.18	0.29
NaHCO₃	S5761	84.020	25.00	2.11
CaCl₂ · 2H₂O	C7902	147.200	1.70	0.25
Na₂-EDTA · 2H₂O	E5134	372.20	0.11	0.04
l-glutamine	G8540	146.10	1.00	0.15
Na-lactate 60% syrup d = 1.32 g/L	L7900	112.100	28.00	3.145.229 g (3.96 mL)
Na-pyruvate	P4562	110.00	0.27	0.03
Glucose	G6152	179.86	5.55	1.00
Penicillin	P4687	–	–	0.05
Streptomycin	S6501	–	–	0.07
BSA		–	–	4.00
CZB-HEPES medium is prepared as for CZB, with the following adjustments: HEPES is added, NaHCO₃ is reduced, and BSA is substituted with PVA.
HEPES-Na (basic)	H3784	260.300	20.00	0.52
NaHCO₃	S5761	84.020	5.00	0.42
PVA	P8136	–	–	0.10
pH 7.4; osmolarity: 266–276 mOsm.
Store for up to 2 wk in refrigerator.